Exclusion of mouse hepatitis virus from a filtered, plastic rodent shipping container during an in transit field challenge.

Although leading suppliers of laboratory mice and rats continue to use filtered shipping boxes to protect their animals from contamination during transport to the end user, no information had been available in the literature to demonstrate that any of these boxes actually accomplish this task. To test this hypothesis, 12 plastic shipping boxes with filters and tight-fitting lids and six cardboard shipping boxes without filters (controls) were each stocked with adult, adventitious disease-free mice. All 18 shipping boxes were transported to a facility housing a breeding colony of mice enzootically infected with four murine viruses, including mouse hepatitis virus (MHV), and were placed inside the colony for 15 h. The boxes were then transported to a commercial testing laboratory, at which the animals were aseptically removed and were held in microisolation cages for 28 days, after which their sera tested for antibody to all four murine viruses. All serum samples from mice held in the control boxes were positive for antibody to MHV, whereas sera from all mice held in filtered boxes were negative for antibody to any of the four viruses. This study demonstrates that at least one type of filtered shipping container protects mice from a field challenge of MHV. To the best of our knowledge, this is the first documentation of any microbial efficacy testing conducted on filtered shipping containers for laboratory animals.

Phylogeny of the defined murine microbiota: altered Schaedler flora.

The "altered Schaedler flora" (ASF) was developed for colonizing germfree rodents with a standardized microbiota. The purpose of this study was to identify each of the eight ASF strains by 16S rRNA sequence analysis. Three strains were previously identified as Lactobacillus acidophilus (strain ASF 360), Lactobacillus salivarius (strain ASF 361), and Bacteroides distasonis (strain ASF 519) based on phenotypic criteria. 16S rRNA analysis indicated that each of the strains differed from its presumptive identity. The 16S rRNA sequence of strain ASF 361 is essentially identical to the 16S rRNA sequences of the type strains of Lactobacillus murinis and Lactobacillus animalis (both isolated from mice), and all of these strains probably belong to a single species. Strain ASF 360 is a novel lactobacillus that clusters with L. acidophilus and Lactobacillus lactis. Strain ASF 519 falls into an unnamed genus containing [Bacteroides] distasonis, [Bacteroides] merdae, [Bacteroides] forsythus, and CDC group DF-3. This unnamed genus is in the Cytophaga-Flavobacterium-Bacteroides phylum and is most closely related to the genus Porphyromonas. The spiral-shaped strain, strain ASF 457, is in the Flexistipes phylum and exhibits sequence identity with rodent isolates of Robertson. The remaining four ASF strains, which are extremely oxygen-sensitive fusiform bacteria, group phylogenetically with the low-G+C-content gram-positive bacteria (Firmicutes, Bacillus-Clostridium group). ASF 356, ASF 492, and ASF 502 fall into Clostridium cluster XIV of Collins et al. Morphologically, ASF 492 resembles members of this cluster, Roseburia cecicola, and Eubacterium plexicaudatum. The 16S rRNA sequence of ASF 492 is identical to that of E. plexicaudatum. Since the type strain and other viable original isolates of E. plexicaudatum have been lost, strain ASF 492 is a candidate for a neotype strain. Strain ASF 500 branches deeply in the low-G+C-content gram-positive phylogenetic tree but is not closely related to any organisms whose 16S rRNA sequences are currently in the GenBank database. The 16S rRNA sequence information determined in the present study should allow rapid identification of ASF strains and should permit detailed analysis of the interactions of ASF organisms during development of intestinal disease in mice that are coinfected with a variety of pathogenic microorganisms.

Compact anaerobic glove box for hospitals and research laboratories.

An anaerobic glove box designed for moderate-sized hospitals or research laboratories is described. The compact vinyl and metal enclosure becomes anaerobic rapidly after inflation, requires no vacuum pump, and provides incubator space equivalent to 15 standard anaerobic jars.

Elimination of the Shigella carrier state in rhesus monkeys (Macaca mulatta) by trimethoprim-sulfamethoxazole.

The Shigella carrier state was eliminated from its nonhuman primate host, Macaca mulatta. Each of 31 animals was treated twice a day for ten consecutive days with 16 mg trimethoprim and 80 mg sulfamethoxazole delivered via stomach tube. Fresh rectal swab and stool enrichment cultures were taken for seven consecutive days as well as the 35th and 78th days after treatment, and all were negative for shigellae. In addition, no clinical signs of shigellosis were observed during or following an extensive period of stress.

Multiple testing for the detection of B virus antibody in specially handled rhesus monkeys after capture from virgin trapping grounds.

Eight groups of rhesus monkeys totaling over 1,000 animals were captured in the virgin trapping grounds of Jammu and Kashmir, India. Individual caging and special handling technics were utilized to prevent cross-contamination during capture, holding, and subsequent shipment to quarantine facilities in the United States. Immediately following the arrival of the monkeys, 5 consecutive blood samples were obtained at approximately 2-wk intervals, and the sera were rested for neutralizing antibody against Herpesvirus simiae. In order to assure the greatest sensitivity possible, sera were not heat-inactivated and were tested against only 10 TCID50 units of virus in addition to the more commonly used concentration of 100 TCID50 units. The first test detected 80-90% of the positive animals within each group, and only 1 seroconversion was noted after the second test. Seventy-three percent of the adults, 36.6% of the young adults, and 12.4% of the juvenile macaques were found to be antibody-positive. Considering the measures employed to prevent cross contamination, these percentages probably reflect the true prevalence of B virus infection in these rhesus monkeys at the time of their capture in the wild.